J_Environmetal Microbiology and Engineering

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Optimal Culture Conditions of Microalgae Haematococcus pluvialis Hyun-Jin PARK 1 , Young-Hwa KIM 2 and Jae-Hwa LEE* 1 1 Department of Bioscience and Biotechnology, College of Medical and Life Science, Silla University, Busan 617-736, Korea. 2 J-20 Department of Pharmaceutical Engineering, College of Medical and Life Science, Silla University, Busan 617-736, Korea. *Corresponding author: phj0386@naver.com The green microalgae Haematococcus pluvialis (H. pluvialis) produces large amounts of astaxanthin. The morphological change of H. pluvialis from green vegetative cells to red resting cyst cells is accompanied by an enhancement in astaxanthin production. To find out high mass culture conditions of H. pluvialis, some important growth factors, such as temperature, initial pH, culture media, CO2, and air effect were tested. The optimal growth condition was found to be as follows: temperature 35℃, initial pH 6.5, 4X MBBM medium. Maximum growth rate of H. pluvialis was obtained at a 5% CO2(0.81 g/L), air(1.06 g/L) environment. From these conditions, it could be possible to cultivate H. pluvialis for large-scale high density. Keywords: optimal culture, Haematococcus pluvialis, marine microalga Heterologous Expression of Metagenome-Derived Wax Ester Synthase in Escherichia coli and Arabidopsis thaliana. Nam-hee KIM 1 , Ji Hye PARK 1 , Eun Sook CHUNG 2 , Jin Cheol KIM 3 , Myung Hwan LEE 1 , Weixin TAO 2 , Eul Chul HWANG 1 , Jai Heon LEE 2 and Seon Woo LEE* 1,2 1 Department of Applied Biology, Dong- A University, Busan 604-714, Republic of Korea. 2 Department of Medical Bioscience, Dong- A University, Busan 604-714, Republic of Korea. 3 J-21 Chemical Biotechnology Research Center, KRICT, Deajeon 305-600, Republic of Korea. *Corresponding author: seonlee@dau.ac.kr We have previously selected a metagenome clone carrying an ORF encoding a wax ester synthase (WES) like protein. Deduced amino acid sequences of the ORF exhibited 30% and 22% identities to WES from Acinetobacter baylyi and Arabidopsis thaliana, respectively. The WES-like protein was functional as a wax ester synthase, when wes gene was expressed in Escherichia coli fed by fatty alcohol, 10 mM of hexadecanol, as a precursor for wax ester synthesis. The unique wax esters produced in E. coli carrying wes gene were identified as wax esters of various chain lengths by GC-MS analysis. The wes gene was also introduced into a model plant, Arabidopsis thaliana, to produce wax esters in a plant system. Transgenic plants carrying the wes gene were obtained by Agrobacterium-mediated transformation. Expression of wes gene in the transgenic plants was confirmed by RT-PCR. Production of wax esters and other phenotypic changes in the transgenic plants are under investigation. 2011 International Symposium & Annual Meeting J-22 Community Structure Analysis of Cyanobacteria using 16S rRNA and Phycocyanin Gene in Korean Reservoirs Kyoung-Hee OH, Seung-Hee SHIN, Eun-Hye PARK, Mi-Na YU and Young-Cheol CHO* Dept. of Environmental Engineering, Chungbuk National University, Cheongju 361-763, Korea. *Corresponding author: choy@cbnu.ac.kr The community structure of cyanobacteria was examined with 16S rRNA and phycocyanin intergenic spacer (PC-IGS) genes as target. Although DGGE analysis with 16S rRNA gene reflected the changes of cyanobacterial community structure with fluctuations of environmental factors, the resolution was restricted to the genus level because of its homogeneity. On the other hand, analysis of PC-IGS gene sequences showed that this gene can classify cyanobacteria in strain level, indicating it is considered as an adequate target gene to analyze the community structure of cyanobacteria in the environmental samples. Moreover, the PC-IGS gene sequences of Microcystis strains in the investigated reservoirs were divided into two clades and each clade exclusively included toxicor non-toxic Microcystis, proposing PC-IGS gene can be used to monitor toxigenic Microcystis at the cyanobacterial bloom events. [Supported by grants from National Science Foundation of Korea] Keywords: Cyanobacteria, PC-IGS, Community structure J-23 Ethanol Fermentation using Main Sugar in Laminaria japonica Geun Hyub KIM, Sung-Mok LEE and Jae-Hwa LEE* Department of Bioscience and Biotechnology, College of Medical and Life Science, Silla University, Busan 617-736, Korea. *Corresponding author: jhalee@silla.ac.kr The excessive use of fossil fuels has recently been causing many problems around the world such as global warming. Therefore, alternative energy source has been of interest, much research as been performed in order to develop alternative energy. This study examined the production of bio-ethanol from the brown-algae main sugar compound. The result of ethanol fermentation used four-bacteria, ethanol production of Zymomonas mobilis KCTC 1534 with different carbon sources (alginate, laminaran, mannitol and Laminaria japonica) confirmed 2.31 g/L, 1.32 g/L and 2.36 g/L respectively. The maximum cell growth with laminaran reached 1.42 (O.D. 600 nm), but other substrate was not significant increases in the fermentation period. The maximum cell growth of Zoogloea ramigera KCCM 40423 was reached approximately 2.61 and 2.37 (O.D. 600 nm) with mannitol and laminaran, respectively. And then ethanol production was 0.97 and 0.236 g/L. Therefore, Laminaran like to be limitations to what ethanol production, but can consume for cell growth increase. Streptomyces venezuelae KCCM 40310, Paracoccous denitrificans KCCM 40267 was not detected ethanol production, but cell growth was significant increases. Keywords: ethanol fermantation, Laminaria japonica, polysaccharides The Korean Society for Microbiology and Biotechnology 401
Identification of Novel Sugar Pathway Genes and their Organization from an Insect Gut Metagenome Chang-Muk LEE* 1 , Sang-Hong YOON 1 , Soo-Jin KIM 2 , So-Hyeon SEO 1 , Young-Seok LEE 1 , Jung-Keun PARK 1 , Han-Chul KANG 1 and Bon-Sung KOO 1 J-24 1 Functional Biomaterial Division, National Academy of Agricultural Science, 2 Rural Development Administration, Suwon 441-707, Korea. National Agrobiodiversity Center, Academy of Agricultural Science, Rural Development Administration, Suwon 441-707, Korea. *Corresponding author: changmuk@rda.go.kr Insect gut symbiotic microbiota play essential roles in the recycling various organic materials, and may thus be exploited as a simple system for processing daily food wastes. To detect the contributing microbial genes, we have constructed 92,544 metagenomic fosmid clones using the larval intestinal microbiome from black soldier fly, Hermetia illucens. The hydrolytic enzymes encoded by uncultured microorganisms were subjected to substrate hydrolysis analysis. Initial screening of the libraries revealed a clone that uses carboxymethyl cellulose as a sole carbon source. Functional screening and metagenome walking with shot-gun libraries revealed two novel gene clusters conferring multiple sugar pathways. The cluster was 36kb in size and transcribed as polycistronic units. They include glycosidase-related sugar pathway genes that are arranged as sugar transferase genes, glycan processing genes, transporter genes, and putative transcription factors. The presence of insertion sequences between clusters suggests that the metagenome-derived clone has acquired the specific organization by lateral gene transfer [Supported by grants from the Rural Development Administration project number PJ0067032011]. Keywords: Metagenome, Hermetia illucens gut, Glycosidase Development of Transgenic Daphnia magna Expressing Enhanced green Fluorescent Protein Thai Hoang LE 1 , Sung-Kyu LEE 2 , Yang-Hoon KIM 3 and Jiho MIN* 1 1 Department of Bioprocess Engineering, Chonbuk National University, 664-14 Duckjin-dong, Jeonju 561-756, South Korea. 2 Toxicology Research Center, Korea Research Institute of Chemical Technology, Daejon, 305-600, Korea. 3 J-25 School of Life Science, Chungbuk National University 410 Sungbong-Ro, Heungduk-Gu, Cheongju 361-763, South Korea. *Corresponding author: jihomin@jbnu.ac.kr To establish a novel system for a rapid detection of environmental stress, a transgenic Daphnia magna able to express a green fluorescent protein has been created. A 32bp DNA fragment containing the 18s-ribosome constitute promoter of daphnia was generated by in-vitro annealing the commercial primers at 80 o C. This target DNA was inserted ahead to the gfp gene in the EGFP vector (Enhanced Green Flourescent Protein). The successful recombinant plasmid confirmed by the DNA sequencing was introduced into the earlier developing-state daphnia eggs (round shape) using microinjection technique. After 48h incubation in the 20 o C chamber, the injected eggs able to develop to juveniles were observed at the 488nm wavelength in the confocal fluorescent microscope to confirm the presence of green fluorescent protein. Our results show that the green fluorescent protein was substantially found in the upper part of the transgenic organisms (e.g., head, back) while no fluorescent signal was detected in the control organism. It demonstrated that a transgenic Daphnia magna expressing green fluorescent protein was firstly generated and would be promising to build a novel testing system for easily monitoring environmental stress. Keywords: Daphnia magna, transgenic, fluorescent 402 www.kormb.or.kr Isolation and Characteristics of Antagonic Microorganisms as Nematicide Against Root-knot Nematode of Fruit Vegetables (Oriental Melon and Cucumber) Beam Soo KIM 1 , Keun-Hyung LEE 1 , Young-Won KANG 1 , Chang-Su KIM 1 , Se-Ho MYEONG 1 , Joung-Ja KWON 2 , Chung-Sig CHOI 3 and Gi-Seok KWON* 1 1 School of Bioresource Sciences, Andong National University, Andong 760-749, Korea. 2 Cheongsong Agricultural Technology Center Cheong Song Gun, CheongSong 763-802, Korea. 3 J-26 Bio Industry Institute, HansBio Co.,Ltd. Andong 760-380, Korea. *Corresponding author: suyaa18@hanmail.net This study was conducted to screen the antagonic microorganisms on root-knot nematode and to utilize in its control. The effect of nematocidal activity of antagonic microorganisms were tested on the nematicide of root-knot nematode, Showing nematocidal activities of 88 strains isolated from various fruit Vegetables field and soil. In vitro study, all tested treatments had nematicidal effect on nematode juveniles 24 and 48 hours from exposures in the 24-well plate. Among them, BS-17, BS-51(2), BS-51(3), BS-52(2), and BS-84(2) were isolated. The highest percentage of nematode mortality was achieved by application of BS-17(96.38%), BS-51(2)(94.11%), BS-51(3)(95.38%), BS-52(2)(94.59%), BS-84(2) (98.48%). Under greenhouse conditions, isolated antagonic microorganisms treatments were effective in reducing population numbers of root-knot nematode and root gall formation in compared to non-treatments. Keywords: Root-knot nematode, Meloidogyne. Antagonic microorganisms, Nematocidal activity, Ne Effect of Plant Growth-Promoting Rhizobacteria on Growth of Chinese cabbage Plant under Salt Stress Hee Sun KOOK 1 , Gun-Woon LEE 1 , Song-Joong YUN 2 , Jong-Chan CHAE 1 and Kui-Jae LEE* 1 J-27 1 Division of Biotechnology, Chonbuk national University, Iksan 570-752, korea. 2 Division of Agriculture Life sciences, Chonbuk National University, Chonju 561-765, Korea. *Corresponding author: k722954@naver.com To gain understanding of responses to salinity stress by rhizobacteria at physiological level, we isolated 15 strains of plant growth-promoting bacteria from soil samples taken in the reclaimed tidelands. These 15 strains have been identified as Pseudomonas, Herbaspirillum, Enterobacter, Serratia, and Stenotrophomonas by 16S rRNA gene sequence analysis. With the exception of Serratia sp., all strains can product indole acetic acid (IAA) and IBA (butyric acid). Among the selected 15 strains, eight strains only showed 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity and rhizobacterial ACC deaminases were detected by polyclonal anti-ACCD antibody. To evaluate whether rhizobacteria inhabiting saline environments confer the resistance against salt stress to Chinese cabbage plant, cabbage seeds were sown in soil inoculated with respective bacterial strains. And then, the early seedling growth was examined under 0 or 100 mM NaCl solution treatment for 2 weeks after germination. Results of our study exhibited that inoculation with selected strains increased the shoot length ranging from 107 to 141%, compared to that of the uninoculated control. These results indicate that the rhizobacterial stains had the potential to promote the growth of cabbage plant under saline condition. Keywords: plant growyh-promoting rhizobacteria (PGPR), Chinese cabbage, NaCl, Salt stress
Page 1 and 2: Isolation and Characterization of L
Page 3 and 4: Characterization of Phosphate-Solub
Page 5: Thermodynamic and Kinetic Analyses
Page 9 and 10: J-32 Enhancement of Biohydrogen Pro
Page 11 and 12: Bioelectrochemical Fixation of CO2
Page 13 and 14: Inhibition of Quorum Sensing-Contro
Page 15 and 16: J-56 Development of Baeyer-Villiger

J_Environmetal Microbiology and Engineering

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